健脾益气方含药血清通过Caspase-3/Vimentin促进人肝癌MHCC-97H细胞凋亡

目的:观察健脾益气方含药血清对人MHCC-97H肝癌细胞凋亡的影响,探讨凋亡过程中波形蛋白(Vimentin)和天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)的变化及其意义。方法:采用血清药理学方法,并结合Vimentin裂解剂和Caspase抑制剂,观察7.5%,15%,30%体积浓度健脾益气方含药血清干预对肝癌细胞增殖、侵袭、凋亡,及Vimentin,Caspase-3,聚腺苷二磷酸核糖聚合酶1(PARP-1)蛋白表达的影响。结果:健脾益气方中、高剂量含药血清均可以降低MHCC-97H肝癌细胞中Vimentin蛋白表达(P0.01),其下调程度与肝癌细胞的增殖抑制率、侵袭抑制率和凋亡率趋势一致;Vimentin裂解组细胞增殖抑制率、侵袭抑制率和凋亡率在各组中均最高;Caspase抑制组细胞的增殖抑制率、侵袭抑制率和凋亡率在各组中均最低。与空白血清组比较,健脾益气方中、高剂量组,Vimentin裂解组Caspase-3蛋白表达上调(P0.01),Vimentin,PARP-1蛋白表达下调(P0.01),且PARP-1蛋白出现了裂解片段;Caspase抑制组Caspase-3表达下调(P0.01),Vimentin,PARP-1蛋白表达无统计学差异,PARP-1无蛋白裂解片段出现。结论:15%的健脾益气方含药血清对人肝癌细胞MHCC-97H的干预效果最佳,可能通过Caspase-3下调细胞中Vimentin蛋白表达,抑制肝癌细胞的增殖与侵袭;同时可能利用Caspase-3/Vimentin形成的正反馈促凋亡信号诱导肝癌细胞发生凋亡。

Effect of Serum Containing Jianpi Yiqi Decoction on MHCC-97H Cell Apoptosis Via Caspase-3/Vimentin

Objective: To investigate the effects of serum containing Jianpi Yiqi decoction (JYD) on MHCC-97H cell apoptosis, and the changes in Vimentin and Caspase-3 in the process of apoptosis and the significance. Method: By using the serologic pharmacological method, Vimentin breaker and Caspase inhibitor were adopted to observe the effect of serum containing Jianpi Yiqi decoction at concentrations of 7.5%, 15% and 30% on proliferation, invasion and apoptosis of liver tumor cells, and protein expressions of Vimentin, Caspase-3 and poly (ADP-ribose) polymerase-1 (PARP-1). Result: Compared with the blank serum group, decreased protein expression of Vimentin and PARP-1(P<0.01), and increased protein expression of Caspase-3 were observed in Jianpi Yiqi decoction groups (with concentrations of 15% and 30%) and Vimentin-breaker group. The down-regulation degree of Vimentin and PARP-1 were consistent with the proliferation inhibition rate, invasion inhibition rate and apoptosis rate of MHCC-97H cells. The proliferation and invasion inhibition rates and apoptosis rate of the Vimentin-breaker group cells were the highest among all experimental groups. However, the Caspase-inhibitor group showed the lowest proliferation and invasion inhibition rates, and apoptosis rate among all experimental groups. Compared with the blank serum group, medium and high-dose JYD group and Caspase-inhibitor group showed up-regulation of Caspase-3 protein expression (P<0.01), down-regulation in Vimentin and PARP-1 expressions (P<0.01), and fragments in PARP-1 protein, the Caspase-inhibitor group showed down-regulation in Caspase-3 expression (P<0.01), with no statistically significant difference in Vimentin and PARP-1 protein expressions. Conclusion: The expression of Vimentin in MHCC-97H cells was down-regulated by serum containing Jianpi Yiqi decoction, especially at the concentration of 15%. Jianpi Yiqi decoction may inhibit liver cancer cell proliferation and invasive by down-regulating Vimentin via Caspase-3, while inducing liver cancer cell apoptosis by using positive feedback apoptosis-promoting apoptosis signal formed by Caspase-3/Vimentin.